Effect of anti-Müllerian hormone in hypothalamic Kiss-1- and GnRH-producing cell models

Abstract

Purpose: Anti-Müllerian hormone (AMH) is one of the local factors involved in follicle development. In addition, AMH and its receptor are broadly expressed throughout the body. In this study, we examined how AMH modifies gene expression of Kiss-1 and GnRH.

Materials and methods: mHypoA-50 and mHypoA-55 cells were originated from the hypothalamic anteroventral periventricular nucleus (AVPV) and arcuate nucleus (ARC), respectively, and these cells are known as Kiss-1 (which encodes kisspeptin) expressing cell models. These cells also express gonadotropin-releasing hormone (GnRH) genes. Our experiments were performed useing these cell models.

Results: Both mHypoA-50 and mHypoA-55 hypothalamic cells expressed AMH and AMH receptor type 2 (AMHR2). Exogenous AMH failed to alter the expression levels of the Kiss-1 gene in both cell models but significantly increased GnRH gene expression by 1.73 ± 0.2-fold at 100 pM in mHypoA-50 AVPV cells and by 1.74 ± 0.17-fold at 1 nM in mHypoA-55 ARC cells. AMH also augmented GnRH protein expression in both cell models. Similar to the phenomenon observed in the hypothalamic cell lines, 100 pM AMH significantly increased GnRH, but not Kiss-1, mRNA expression in primary cultures of fetal rat brain cells. Kisspeptin-10 (KP10) increased Kiss-1 gene expression in mHypoA-55 ARC cells but this was blocked by AMH. AMH did not alter the expression of the kisspeptin receptor (Kiss1R) or that of neurokinin B or dynorphin A in mHypoA-55 ARC cells.

Conclusions: It was demonstrated that AMH participates in hypothalamic-pituitary-gonadal axis control by stimulating GnRH expression. In addition, AMH might be a potent repressor of Kiss-1 gene expression induced by KP10.

抗缪勒管激素在下丘脑Kiss-1和促性腺激素释放激素生成细胞模型中的作用 摘要

目的：抗缪勒管激素 (AMH) 是参与卵泡发育的局部因素之一。此外, AMH 及其受体在全身广泛表达。在这项研究中, 我们研究了AMH如何修饰 Kiss-1 和 GnRH 的基因表达。

材料和方法：mHypoA-50 和 mHypoA-55 细胞分别来源于下丘脑前腹侧周围核团 (AVPV) 和弓状核 (ARC), 这些细胞被称为 Kiss-1（编码kisspeptin）表达细胞模型。这些细胞还表达促性腺激素释放激素 (GnRH) 基因。我们的实验是应用这些细胞模型进行的。

结果：mHypoA-50和mHypoA-55下丘脑细胞均表达AMH和AMH受体2型（AMHR2）。

100pM外源性AMH并不改变两种细胞模型中Kiss-1基因的表达水平, 但mHypoA-50 AVPV细胞中GnRH 基因表达显著增加1.73±0.2倍, 而1 nM AMH可使 mHypoA-55 ARC细胞中GnRH基因表达增加了1.74±0.17倍。AMH 还促进了这两种细胞模型中的GnRH蛋白表达。与在下丘脑细胞系中观察到的现象类似, 在胎鼠脑原代细胞中100 pM AMH显著增加 GnRH的mRNA表达, 但不增加 Kiss-1的mRNA表达。Kisspeptin-10 (KP10) 增加了 mHypoA-55 ARC细胞中Kiss-1基因的表达, 而AMH抑制表达。AMH不会改变 mHypoA-55 ARC细胞中Kisspeptin受体 (Kiss1R)、神经激肽B、强啡肽A的表达。

结论：已证明AMH通过激发 GnRH表达参与调控下丘脑-垂体-性腺轴。此外, AMH可能是KP10诱导的Kiss-1基因表达的有效阻遏物。

Keywords:

• Anti-Müllerian hormone
• kisspeptin
• gonadotropin-releasing hormone

Acknowledgments

The manuscript was edited by ThinkSCIENCE, Inc., Tokyo, Japan.

Disclosure statement

The authors declare they have no competing interests.

Additional information

Funding

This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (HK and AO).